Somatic embryogenesis and

synthetic seed production

Gaurav Chandra Gyanwali
M.Sc Biotechnology
TU, Nepal

What are synthetic seeds?

Synthetic seeds are encapsulated somatic

embryos
The seeds may be dry or hydrated. Dry
seeds are either naked or encapsulated in
polyoxyethylene glycol (Polyox) followed by

What are synthetic seeds?

Alginate encapsulated synthetic seeds (left) and synthetic seed derived plantlets
in orchids (Saiprasad, 2001)

Somatic embryogenesis
It is the process of production of embryos
from somatic cells (not as a result of
gametic fusion)
The somatic embryos are commonly
called embryoids or non-zygotic embryos
Can be induced directly or indirectly
Somatic embryogenesis proceeds in two
steps: induction and maturation
Induction requires high auxin and low
NH4+, while maturation requires low or no
auxin and high sucrose (6-40%)
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Somatic embryogenesis../2
derived from single cell
which
divide
to
form
meristemoid

Cells
of
meristemoid
contain dense cytoplasm,
large starch grains, large
nucleus and nucleolus with
high dehydrogenase activity
These cells later become
cutinised
and
lose
cytoplasmic connection with
the
surroundingthe
proembryo stage
The proembryo first forms
globular structure and then
proceeds to form heart and
torpedo
embryoids,
respectively
In the last step, the
embryoids
develop
into

Production of synthetic seeds

Production of synthetic seeds

from somatic embryos
In order to use somatic embryos for
artificial seed production, they should be
optimized.
Then there should be the synchronization
of somatic embryo production
The somatic embryos thus produced
should then be separated and subjected to
mass scale production of somatic embryo

They
are
then
subjected
to
standardization of encapsulation and
artificial endosperms

Optimization of somatic embryo

production
Should be done on species specific, explants
specific manner
There is no sure shot mechanism, optimal
conditions should be found out empirically trying
out different culture conditions, hormonal
regimes, media, etc
Conditions optimal for one explants may not suit
that for other explants even from the same
species
The embryos thus produced should be vigorous
and of high quality such that they can produce
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plants in frequencies comparable to natural

Optimization of gelling agents

and encapsulation of somatic
embryos

The gelling agents used should have low

toxicity to somatic embryos, should be
cheap, rapidly gelling and should have
moderate viscosity.
Several gelling agents like agar, alginate,
carboxy methyl cellulose, carrageenan,
sodium pectate, etc. have been tested
Alginate has so far found to be the best
due to its enhanced encapsulation and
rigidity of the capsules against mechanical
injury
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Optimization of gelling agents

and encapsulation of somatic
embryos../2

Somatic embryos are mixed with sodium

alginate gel (0.5 5% w/v) and dropped
into calcium salt solution (CaCl2 (30100
mM), Ca(NO3)2 (30100 mM)).
As a result of ion-exchange reaction,
sodium ions are replaced by calcium ions
forming calcium alginate beads or
capsules
surrounding
the
somatic
embryos.
The size of the capsule is controlled by
varying the inner diameter of the pipette
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nozzle.

Optimization of gelling agents

and encapsulation of somatic
embryos../3
Hardening of the calcium alginate is
modulated with the concentrations of
sodium alginate and calcium chloride as
well as the duration of complex formation.
The use of a dual nozzle pipette in which
the embryos flow through the inner
pipette and the alginate solution through
the outer pipette is suitable for
encapsulation
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Optimization of artificial
endosperm
Somatic embryos lack true endosperm.
This has been a major limitation in
imposing dormancy and stress tolerance
in artificial seeds
So they need to be supplied with artificial
sources of nourishment and growth
regulators, but the protocols are not well
established as yet.

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Advantages of using synthetic

seeds
Ease of handling (during
production/storage)
Easy to transport
Potential for long term storage without
loss of viability
Maintain the clonal nature (i.e., genetic
uniformity) of the resulting plants
Serves as a channel for new plant lines
produced through biotechnological
advances
directly to the greenhouse or field
Allows economical mass propagation of
elite plant varieties.

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Limitations
Limited production of viable
micropropagules useful in synthetic seed
production.
Anomalous and asynchronous development
of somatic embryos.
Improper maturation of the somatic
embryos that makes them inefficient for
germination and conversion into normal
plants.
Lack of dormancy and stress tolerance in
somatic embryos that limit the storage of
synthetic seeds.
Poor conversion of even apparently normally
matured somatic embryos and other
micropropagules into plantlets that limit the 15
value of the synthetic seeds and ultimately